Daniel:Notebook/ComboLock/2017-4-12: Difference between revisions
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{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | ||
|- style="font-size:12pt;font-weight:bold" align="center" | |- style="font-size:12pt;font-weight:bold" align="center" | ||
|style="background-color:#8DB4E2" width=" | |style="background-color:#8DB4E2" width="230" height="30" | Reagent | ||
|style="background-color:#8DB4E2" width="85" | Single Rxn Vol | |style="background-color:#8DB4E2" width="85" | Single Rxn Vol | ||
|style="background-color:#95B3D7" width="85" | Master Mix (13.2X) | |style="background-color:#95B3D7" width="85" | Master Mix (13.2X) | ||
Line 18: | Line 18: | ||
|- style="font-size:12pt" | |- style="font-size:12pt" | ||
| height="15" valign="bottom" | RCA Rxn | | height="15" valign="bottom" | RCA Rxn | ||
| align="center" align="center" valign="bottom" | | | align="center" align="center" valign="bottom" | 7 | ||
| align="center" align="center" valign="bottom" | 0 | | align="center" align="center" valign="bottom" | 0 | ||
|- style="background-color:#BFBFBF;font-size:12pt" | |- style="background-color:#BFBFBF;font-size:12pt" | ||
| height="15" valign="bottom" | | | height="15" valign="bottom" | 10X Buffer 3.1 | ||
| align="center" align="center" valign="bottom" | 2 | | align="center" align="center" valign="bottom" | 2 | ||
| align="center" align="center" valign="bottom" | 26.4 | | align="center" align="center" valign="bottom" | 26.4 | ||
|- style="font-size:12pt" | |- style="font-size:12pt" | ||
| height="15" valign="bottom" | Bglii | | height="15" valign="bottom" | Bglii | ||
| align="center" align="center" valign="bottom" | 1 | | align="center" align="center" valign="bottom" | 1 | ||
| align="center" align="center" valign="bottom" | 0 | | align="center" align="center" valign="bottom" | 0 | ||
|- style="font-size:12pt" | |- style="background-color:#BFBFBF;font-size:12pt" | ||
| height="15" valign="bottom" | nfH2O | | height="15" valign="bottom" | nfH2O | ||
| align="center" align="center" valign="bottom" | 10 | | align="center" align="center" valign="bottom" | 10 | ||
| align="center" align="center" valign="bottom" | 132 | | align="center" align="center" valign="bottom" | 132 | ||
|- style=" | |- style="font-size:12pt" | ||
| height="15" valign="bottom" | Total | | height="15" valign="bottom" | Total | ||
|style="font-weight:bold" align="center" align="center" valign="bottom" | 20 | |style="font-weight:bold" align="center" align="center" valign="bottom" | 20 | ||
|style="font-weight:bold" align="center" align="center" valign="bottom" | | |style="font-weight:bold" align="center" align="center" valign="bottom" | 158.4 | ||
|} | |} | ||
<li>Aliquot | <li>Aliquot 12 uL master mix into new tube</li> | ||
<li>Add | <li>Add 7 uL appropriate sample</li> | ||
<li>Use the following thermocycler program</li> | <li>Use the following thermocycler program</li> | ||
<ol type="a"> | <ol type="a"> |
Revision as of 15:13, 12 April 2017
RCA Double Deuce
Protocol
- Bglii digestion
- Make the following reactions; Do not add Bglii yet
- Aliquot 12 uL master mix into new tube
- Add 7 uL appropriate sample
- Use the following thermocycler program
- 2 hr 37C; When this cycle starts add 1 uL Bglii
- Heat kill with 65C for 20 min
Reagent | Single Rxn Vol | Master Mix (13.2X) |
RCA Rxn | 7 | 0 |
10X Buffer 3.1 | 2 | 26.4 |
Bglii | 1 | 0 |
nfH2O | 10 | 132 |
Total | 20 | 158.4 |